大豆原生质体愈伤组织的体细胞胚胎发生(英文)  

Somatic Embryogenesis from Protoplast-derived Callus in Soybean ( Glycine max L. )

作　　者：成雄鹰[1] 

机构地区：[1]浙江农业大学原子核农业科学研究所,杭州310029

出　　处：《浙江农业大学学报》1991年第1期103-110,共8页

摘　　要：在改良MS培养基上从5个大豆品种成熟种子来源的幼苗未展开叶片上诱导出愈伤组织,利用2个月月龄的这些愈伤组织在摇床上建立了细胞悬浮培养系,细胞悬浮培养系每1至2周转培1次,并用于原生质体分离,转培后3到5天的细胞经酶解后原生质体的产量最高,纯化后的原生质体用3种方法进行培养,即液体培养法、琼脂包埋法和琼脂底层法,在琼脂底层法中,培养皿底层先浇上无甘露醇的琼脂培养基,而原生质体则悬浮在2到4毫升含0.45 mol甘露醇的培养基中,3种方法以琼脂底层法最佳,原生质体在培养后2到3天内能见到第1次细胞分裂,在适当的条件下1周内高达80％的原生质体发生分裂,置板频率一般在40％到60％之间,5个供试品种中4个的原生质体愈伤组织在固体和液体培养中出现体细胞胚胎发生,在液体培养条件下得到了大量子叶和胚根发育良好的体细胞胚,但由于这些胚未进一步萌发,故未能得到完整的再生植株。Calli induced from the unexpanded leaves of mature seed-derived seedlings of five soybean cultivars on modified MS medium were suspended in liquid medium on a shaker at 90 rpm after two-month solid culture.The suspension cultures were diluted every 1 to 2 weeks and were used as sources for protoplast isolation.The highest yields of protoplasts were obtained when the cells were treated with 0.1~0.29% pectolyase Y-23 and cellulase 'Onozoka R-10' or 'RS' in 0.45 mol D-mannitol solutions 3~5 days after subculture.Purified protoplasts were cultured in three ways,liquid culture,solidified culture and bottom agar culture.Best results were obtained in bottom agar culture where protoplasts were suspended in 2~3 ml high osmotic liquid medium contained in7plastic dishes already poured with the same or different agar medium without mannitol.The first cell divisions were observed 2~3 days after culture and as high as 80% cultured protoplasts underwent cell divisions within a week after culture in some cases and plating efficiency varied from 40% to 60% Visible protocolones were transferred to various solidified and liquid media for embryogenesis and plant regeneration.Somatic embryos were induced in four of the five cultivars in modified MS medium containing 2 mg/1 2,4-D.Plant regeneration from these protoplast-derived embryos was unsuccessful due to inability of the embryos to germinate.

关 键 词：大豆 细胞悬浮 培养 原生质 体细胞 

分 类 号：S565.103.5[农业科学—作物学]